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. 2021 Oct 18;11:20627. doi: 10.1038/s41598-021-99974-7

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Detailed analysis of nematocyst discharge inhibition for three candidate molecules. (a) Structures of compounds 1, 2, and 3. (b) Prey capture inhibition at different concentrations of selected small molecules. Animals were treated in triplicates with the indicated concentration of the compounds for 30 min. Nematocyst discharge was recorded by counting the fraction of artemia (out of ten) caught by each polyp. The inhibitor concentration is shown on the log scale. Colored areas correspond to standard deviation. IC50 values were determined by fitting sigmoid curves to the average inhibition (dashed lines) and solving them for 50% inhibition. (c) Reversibility assay of mechanosensitive nematocyst discharge was tested by recording discharge in the presence of each selected compound at 12.5 µM and again immediately after compound removal (wash). Nematocyst discharge was evaluated by counting the number of attached stenoteles after mechanical stimulus with a gelatin-covered fishing line. Error bars represent standard deviations from triplicate experiments. STM, Streptomycin. (d) Immunostaining of Hydra whole-mounts with a pan-TRPN antibody shows a prominent signal at the base of cnidocils (white arrows) extending from battery cells of the tentacle surface. Control animals were treated with DMSO. (e) Treatment with compound 2 for 2 h leads to a loss of the signal probably due to the relocalization of TRPN in the battery cells. Scale bars = 10 µm. The insets show exemplary cnidocils at higher magnification. Images were acquired with a Nikon A1 confocal microscope using imaging software NIS Elements (AR 45.51.01).